Publication: Purification and characterization of a Bacillus circulans No. 4.1 chitinase expressed in Escherichia coli
2
Issued Date
2006-04-01
Resource Type
ISSN
09593993
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2-s2.0-33646442183
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Mahidol University
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SCOPUS
Bibliographic Citation
World Journal of Microbiology and Biotechnology. Vol.22, No.4 (2006), 331-335
Suggested Citation
Patcharaporn Siwayaprahm, Mongkon Audtho, Kunio Ohmiya, Chanpen Wiwat Purification and characterization of a Bacillus circulans No. 4.1 chitinase expressed in Escherichia coli. World Journal of Microbiology and Biotechnology. Vol.22, No.4 (2006), 331-335. doi:10.1007/s11274-005-9038-8 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/23057
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Title
Purification and characterization of a Bacillus circulans No. 4.1 chitinase expressed in Escherichia coli
Abstract
A DNA fragment encoding for 598 amino acids of chitinase protein from Bacillus circulans No. 4.1 was subcloned into pQE-30 expression vector and transformed into Escherichia coli M15 (pREP4). The molecular weight of the expressed protein was approximately 66 kDa. Enzymatic activity of the recombinant protein was assayed after purification using affinity chromatography on a nickel chelating resin. The enzyme hydrolyzed N- acetylchitooligosaccharides mainly to N-acetylchitobiose, and was active toward chitin, carboxymethyl-chitin, colloidal chitin, glycol chitin and 4-methylumbelliferyl-β-D-N, N′-diacetylchitobiose. The pH and temperature optima of the chitinase enzyme were 7.0 and 45°C, respectively. This enzyme was stable in the pH range of 5.0-9.0 and at temperatures up to 50°C. In addition, when cleaved by a proteolytic enzyme, the 20-kDa product could retain high chitinolytic activity. © Springer 2005.